Novel traf6 inhibiting protein

Abstract

Disclosed are novel isolated tumor necrosis factor receptor associated factor (TRAF) 6 inhibiting proteins or functional derivatives thereof The proteins contain a KRAB domain at their N-termini and 14 C 2 H 2 type zinc finger motifs at their C-termini, bind to TFAF6 but do not bind to TRAF2 and TRAF3, and inhibit activities of NF-κB or AP-1 in a signal transduction pathway of tumor necrosis factor (TNF) by interacting with TRAF6.

What is claimed is:  
     
         1 . An isolated TRAF6-inhibiting protein comprising a KRAB domain at its N-terminus and fourteen C2H2 type zinc finger motifs at its C-terminus, which binds to TRAF6 but does not bind to TRAF2 and TRAF3, and inhibits activity of TRAF6, NF-κB or AP-1.  
     
     
         2 . The isolated TRAF6-inhibiting protein of  claim 1 , wherein the TRAF6-inhibiting protein is derived from human.  
     
     
         3 . The isolated TRAF6-inhibiting protein of  claim 1 , wherein the TRAF6-inhibiting protein comprises the sequence described in SEQ ID NO: 7.  
     
     
         4 . A nucleic acid sequence encoding the TRAF6-inhibiting protein defined in any of  claims 1  to  3 .  
     
     
         5 . The nucleic acid sequence of  claim 4 , wherein the nucleic acid sequence comprises gDNA, cDAN or RNA.  
     
     
         6 . A nucleic acid sequence comprising the sequence described in SEQ ID NO: 6.  
     
     
         7 . A vector comprising a nucleic acid sequence encoding the TRAF6-inhibiting protein defined in  claim 1 .  
     
     
         8 . The vector of  claim 7 , wherein the nucleic acid sequence comprises the sequence described in SEQ ID NO: 6.  
     
     
         9 . A host cell or cell line transformed or transfected with the vector defined in claims  7  or  8 .  
     
     
         10 . The transfected host cell or cell line of  claim 9 , wherein the host cell is yeast, 293T cell or 293/EBNA.  
     
     
         11 . A method for producing isolated TRAF6-inhibiting proteins comprising culturing the transformed or transfected host cell or cell line of  claim 9  and purifying TRAF6-inhibiting proteins from the culture.  
     
     
         12 . A method for purifying a molecule capable of selectively binding to the TRAF6-inhibiting proteins of  claim 1  comprising culturing the TRAF6-inhibiting protein with a candidate molecule, isolating the resulting complex, and dissociating the TRAF6-inhibiting protein from the isolated complex.  
     
     
         13 . The method of  claim 12 , wherein the candidate molecule is TRAF6.  
     
     
         14 . A method for screening a substance capable of modulating activity of a TRAF6-inhibiting protein comprising culturing a mixture containing TRAF6 and TRAF6-inhibiting protein along with a candidate molecule and detecting the ability of the candidate molecule to modulate the formation of the TRAF6/TRAF6-inhibiting protein complex or the ability of the candidate molecule to prevent or inhibit the dissociation of the TRAF6/TRAF6-inhibiting protein complex.  
     
     
         15 . An antibody directed against the TRAF6-inhibiting protein of  claim 1 .  
     
     
         16 . A pharmaceutical composition useful in treating or preventing diseases associated with hyperactivity of TRAF6, NF-κB or AP- 1, comprising a therapeutically or prophylactically effective amount of an active ingredient selected from the group consisting of expression vectors of TRAF6-inhibiting proteins, isolated mRNAs of TRAF6-inhibiting proteins, TRAF6-inhibiting proteins, and mixtures of any two or more thereof, alone or in combination with any pharmaceutically acceptable carrier.  
     
     
         17 . The pharmaceutical composition of  claim 16 , wherein the disease is selected from a group consisting of osteoporosis, atherosclerosis, asthma, arthritis, cachexia, cancer, diabets, inflammatory bowel diseases, stroke and septic shock.  
     
     
         18 . A method for treating or preventing diseases associated with hyperactivity of TRAF6, NF-κB or AP- 1, comprising administering a composition comprising a therapeutically or prophylactically effective amount of an active ingredient selected from the group consisting of expression vectors of TRAF6-inhibiting proteins, isolated mRNAs of TRAF6inhibiting proteins, TRAF6- biting proteins, and mixtures of any two or more thereof alone or in combination with any pharmaceutically acceptable carrier, to an animal.  
     
     
         19 . A phannaceutical composition useful in treating or preventing diseases associated with hypoactivity of TRAF6, NF-κB or AP- 1, comprising a therapeutically or prophylactically effective amount of an active ingredient selected from the group consisting of antisense RNAs of TRAF6-inhibiting proteins, antibodies directed against TRAF6-inhibiting proteins, and mixtures of any two or more thereof, alone or in combination with any pharmaceutically acceptable carrier.  
     
     
         20 . The pharmaceutical composition of  claim 19 , wherein the disease is selected from a group consisting of AIDS, neurodegenerative disorders, autoimmune diseases, immunodeficiencies and stroke.  
     
     
         21 . A method for treating or preventing diseases associated with hypoactivity of TRAF6, NF-κB or AP-1, comprising administering a composition comprising a therapeutically or prophylactically effective amount of an active ingredient selected from the group consisting of antisense RNA of TRAF6-inhibiting proteins, antibodies directed against TRAF6-inhibiting proteins, and mixtures of any two or more thereof alone or in combination with any pharmaceutically acceptable carrier, to an animal.  
     
     
         22 . A recombinant plasmid deposited as a Deposit Accession No. KCTC-0963BP.  
     
     
         23 . A pharmaceutical composition for treating or preventing metabolic bone diseases comprising an active ingredient selected from a group consisting of expression plasmids of TRAF6-inhibiting proteins, isolated mRNAs of TRAF6-inhibiting proteins, TRAF6-inhibiting proteins, and mixtures of any two or more thereof, alone or in combination with any pharmaceutically acceptable carrier.  
     
     
         24 . The pharmaceutical composition of  claim 23 , wherein the metabolic bone disease is selected from a group consisting of osteoporosis, lesions on the bone induced by bone metastasis of tumors, bone cancers of primary type, rheumatoid or degenerative arthritis, periodontal disease, inflammatory alveolar bone resorption diseases arising after implantation of a dental implant inflammatory bone resorption disease caused by implantation for fixing bone and Paget's disease.  
     
     
         25 . A method for treating or preventing metabolic bone diseases, comprising administering a composition comprising an active ingredient selected from a group consisting of expression plasmids of TRAF6-inhibiting proteins, isolated mRNAs of TRAF6-inhibiting proteins, TRAF6-inhibiting proteins, and mixtures of any two or more thereof, alone or in combination with any pharmaceutically acceptable carrier, to an animal.